|本期目录/Table of Contents|

[1]马梦楚,任媛媛,刘欣.肺腺癌细胞中PRDM5抑癌作用的分子机制初探[J].天津医科大学学报,2022,28(04):360-365.
 MA Meng-chu,REN Yuan-yuan,LIU Xin.Preliminary study on the molecular mechanism of PRDM5 suppression in lung adenocarcinoma[J].Journal of Tianjin Medical University,2022,28(04):360-365.
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肺腺癌细胞中PRDM5抑癌作用的分子机制初探(PDF)
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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:
28卷
期数:
2022年04期
页码:
360-365
栏目:
基础医学
出版日期:
2022-07-20

文章信息/Info

Title:
Preliminary study on the molecular mechanism of PRDM5 suppression in lung adenocarcinoma
文章编号:
1006-8147(2022)04-0360-06
作者:
马梦楚任媛媛刘欣
(天津医科大学基础医学院生物化学与分子生物学系,天津300070)
Author(s):
MA Meng-chuREN Yuan-yuanLIU Xin
(Department of Biochemistry and Molecular Biology,School of Basic Medical Sciences,Tianjin Medical University,Tianjin 300070,China)
关键词:
PRDM5NuRD复合物免疫共沉淀肺腺癌
Keywords:
PRDM5NuRD complexCo-IPlung adenocarcinoma
分类号:
Q786
DOI:
-
文献标志码:
A
摘要:
目的:解析肺腺癌中PR结构域锌指蛋白5(PRDM5)发挥抑癌功能的分子机制,寻找与PRDM5相互作用的蛋白因子。方法:免疫共沉淀实验(Co-IP)检测肺腺癌细胞系A549和H1299中PRDM5与核小体重塑和去乙酰化酶复合物(NuRD复合物)各组分之间内源性结合。构建含有GST标签的原核表达载体pGEX-4T-1-HDAC1、pGEX-4T-1-MTA1、pGEX-4T-1-MTA2以及带有His标签的原核表达载体pET-28a(+)-PRDM5。将上述重组质粒转化入大肠埃希菌(E.coli)BL21中诱导融合蛋白成功表达并纯化相应的蛋白。GST-pull down体外实验检测PRDM5与NuRD复合物直接结合的组分。结果:在肺腺癌细胞系A549和H1299中证实PRDM5与NuRD复合物多个主要组分CHD3、CHD4、MTA1/2、HDAC1/2、RbAp46/48、MBD2/3间存在内源性结合。体外GST-pull down实验证实PRDM5与NuRD复合物中的组蛋白去乙酰化酶1(HDAC1)、转移相关蛋白1(MTA1)和转移相关蛋白2(MTA2)组分存在直接结合。结论:肺腺癌细胞中PRDM5与NuRD复合物存在相互作用。
Abstract:
Objective: To elucidate the molecular mechanism of PR/SET Domain zinc finger protein 5(PRDM5) suppression in lung adenocarcinoma,and to find the protein factors interacting with PRDM5. Methods: Co-IP assay was used to detect endogenous binding between PRDM5 and Nucleosome Remodeling and histone Deacetylation complex(NuRD complex) components in lung adenocarcinoma cell lines A549 and H1299. GST-tagged prokaryotic expression vectors pGEX-4T-1-HDAC1,pGEX-4T-1-MTA1,pGEX-4T-1-MTA2 and His-tagged pET-28a(+)-PRDM5 were constructed. Recombinant plasmids were transformed into E.coli BL21 to induce the expression of the fusion proteins and purify the corresponding protein,respectively. The components of NuRD complex which directly bind with PRDM5 were determined by GST-pull down. Results: Most components of NuRD complex can bind with PRDM5 endogenously in lung adenocarcinoma cells A549 and H1299,such as CHD3,CHD4,MTA1/2,HDAC1/2,RbAp46/48 and MBD2/3. Direct binding of PRDM5 with histone deacetylase 1(HDAC1),metastasis associated protein 1(MTA1) and metastasis associated protein 2(MTA2) components in NuRD complex were confirmed by GST-pull down in vitro. Conclusion: PRDM5 interacts with NuRD complex in lung adenocarcinoma cells.

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备注/Memo

备注/Memo:
基金项目 国家自然科学基金面上项目(81572867);天津市教委科研计划项目(自然科学)(2019KJ171)
作者简介:马梦楚(1998-),女,硕士在读,研究方向:抑癌因子PRDM5在肿瘤发生发展中的作用;通信作者:刘欣,E-mail:xinliu_0828@tmu.edu.cn。
更新日期/Last Update: 2022-07-20